Clinical trials are an important method for evaluating the safety and efficacy of in vitro diagnostic reagents, and are a key basis for product registration review and approval. In order to strengthen the management of clinical trials of in vitro diagnostic reagents, the National Medical Products Administration and relevant departments have formulated a series of regulations at the regulatory level, and require applicants and clinical trial institutions to establish a quality management system for clinical trials of in vitro diagnostic reagents. Medical laboratory is the main department and implementer of in vitro diagnostic reagent clinical trials in medical institutions. In recent years, with the rapid development of the in vitro diagnostic industry, the clinical trial projects of in vitro diagnostic reagents conducted by medical laboratory have been increasing day by day. However, there are currently few discussions on the clinical trial of in vitro diagnostic reagents from the perspective of researchers. Therefore, this article summarizes the characteristics of clinical trials of in vitro diagnostic reagents, analyzes the problems and difficulties in conducting clinical trials of in vitro diagnostic reagents in current medical laboratories, and introduces the laboratory′s experience in management; to provide reference for medical testing laboratories that have not yet conducted or have already conducted clinical trials of in vitro diagnostic reagents, in order to improve the quality and efficiency of clinical trials.

Clinical trials are an important method for evaluating the safety and efficacy of in vitro diagnostic reagents, and are a key basis for product registration review and approval. In order to strengthen the management of clinical trials of in vitro diagnostic reagents, the National Medical Products Administration and relevant departments have formulated a series of regulations at the regulatory level, and require applicants and clinical trial institutions to establish a quality management system for clinical trials of in vitro diagnostic reagents. Medical laboratory is the main department and implementer of in vitro diagnostic reagent clinical trials in medical institutions. In recent years, with the rapid development of the in vitro diagnostic industry, the clinical trial projects of in vitro diagnostic reagents conducted by medical laboratory have been increasing day by day. However, there are currently few discussions on the clinical trial of in vitro diagnostic reagents from the perspective of researchers. Therefore, this article summarizes the characteristics of clinical trials of in vitro diagnostic reagents, analyzes the problems and difficulties in conducting clinical trials of in vitro diagnostic reagents in current medical laboratories, and introduces the laboratory′s experience in management; to provide reference for medical testing laboratories that have not yet conducted or have already conducted clinical trials of in vitro diagnostic reagents, in order to improve the quality and efficiency of clinical trials.

Objective·To study the relationship between evolution and the developmental process from the perspective of DNA sequence conservation,and explore their inherent principles.Methods·First,conservation rate(CR)was established by analyzing the conservation of amino acid sequences of coding genes in 100 species to quantify the evolutionary conservation of genes.The relationship between CR and developmental potential was verified by using the feature genes involved in embryonic stem cells pathways.Secondly,cell type-specific genes and their characteristics in conservation were studied by analyzing the RNA sequencing(RNA-seq)data of the three early germ layers(ectoderm,mesoderm and endoderm)and their corresponding mature organs(brain,heart,liver,etc).Then,chromatin immunoprecipitation sequencing(ChIP-seq)data of enhancer histone H3 acetylated at lysine 27(H3K27ac)from early germ layers and mature organs were collected to search for enhancer sites and identify super enhancers in various cells and tissues by using the ROSE procedure.Functional enrichment and signaling pathway analysis of genes was used to examine the identity correlation between SEs-regulated genes and the corresponding cell characteristics,to clarify whether the SEs identified in this study were consistent with the characteristics reported in previous studies.Finally,PhastCons program was used to calculate the DNA conservation score(CS)of non-coding regulatory regions to study their relationship with developmental potential.Results·In the coding region of DNA,CR was successfully established to quantify the conservation of genes.The gene expression data of early germ layers and mature organs showed that the genes with higher conservation rate were more relevant to the stemness and early developmental process,and the differences between the tissues from early and late development could be distinguished by using CR.In the non-coding regions of DNA,it was found that the conservation of regulatory regions was also correlated with development.The CS of the SE sequences in the early developmental germ layers was significantly higher than that of the SE sequences in the corresponding mature organs.However,cell-specific typical enhancers(TEs)did not show such a trend.Conclusion·During the developmental process,CR of genes expressed in the coding region decreases,and CS of super-enhancer DNA in the non-coding region decreases.

The importance of structural variants(SVs)for human phenotypes and diseases is now recognized.Although a variety of SV detection platforms and strategies that vary in sensitivity and specificity have been developed,few benchmarking procedures are available to confidently assess their performances in biological and clinical research.To facilitate the validation and application of these SV detection approaches,we established an Asian reference material by characterizing the genome of an Epstein-Barr virus(EBV)-immortalized B lymphocyte line along with identified benchmark regions and high-confidence SV calls.We established a high-confidence SV callset with 8938 SVs by integrating four alignment-based SV callers,including 109x Pacific Biosciences(PacBio)continuous long reads(CLRs),22 x PacBio circular consensus sequencing(CCS)reads,104x Oxford Nanopore Technologies(ONT)long reads,and 114×Bionano optical mapping plat-form,and one de novo assembly-based SV caller using CCS reads.A total of 544 randomly selected SVs were validated by PCR amplification and Sanger sequencing,demonstrating the robustness of our SV calls.Combining trio-binning-based haplotype assemblies,we established an SV benchmark for identifying false negatives and false positives by constructing the continuous high-confidence regions(CHCRs),which covered 1.46 gigabase pairs(Gb)and 6882 SVs supported by at least one diploid haplotype assembly.Establishing high-confidence SV calls for a benchmark sample that has been characterized by multiple technologies provides a valuable resource for investigating SVs in human biology,disease,and clinical research.

Objective:To analyze the clinical effect of hyaluronidase injection through the facial artery in the treatment of vascular embolism such as skin ulceration and necrosis after cosmetic injection.Methods:Hyaluronidase was injected through facial artery in 13 patients who were diagnosed with vascular embolism after facial injection from January 2019 to May 2020. The facial artery was punctured with 22-gauge arterial blood collection needle or 19/23-gauge disposable venous infusion needle. The angle between the needle body and the skin varies depending on the patients’ weight, ranged 30°-45°. The needle was advanced slowly and pushed forward by 2-3 mm when blood backflow appeared in the needle core. After confirming the successful puncture of the facial artery, 0.5-1.5 ml hyaluronidase was slowly injected into the facial artery. The time of skin relaxation, tenderness relief, ulcer healing and wound recovery were observed. The pigmentation was observed and the Vancouver Scar Scale (VSS) was used to score the scars after 3-12 months.Results:A toal of 13 patients with vascular complications of hyaluronidase filler were retrospectively reviewed. The patients were 18-45-year-old(mean age, 35 years) and received hyaluronidase filler at private clinics. There were 12 women and 1 man. The time from onset to visit was 14 h to 4 d, with an average time of 2.5 d. Hyaluronidase was most commonly injected into the nasolabial folds (54%, 7 of 13). The second-ranked area is the nasalroot (23%, 3 of 13). These patients had skin swelling, necrosis, ecchymosis or black scabs during or after hyaluronidase injection. Some patients showed skin lesions combined with oral ulcer. After percutaneous facial arterial hyaluronidase injection, the local skin tissue injuries of the 13 patients were improved in time. The time of skin relaxation was (0.77±0.25) d, the time of tenderness relief was (1.23±0.64) d, the time of ulcer healing was (3.14±0.64) d and the time of wound recovery was (5.85±0.86) d. Patients were followed up for 3-12 months, with an average of 7 months. One patient had slight scar (VSS score of 1), two patients had only mild pigmentation (VSS score of 0), and the other ten patients had no scar and pigmentation (VSS score of 0).Conclusions:It is effective to improve local microcirculation and reduce skin tissue injury after percutaneous facial artery hyaluronidase injection in the treatment of skin injury caused by facial filler injection.

Objective:To analyze the clinical effect of hyaluronidase injection through the facial artery in the treatment of vascular embolism such as skin ulceration and necrosis after cosmetic injection.Methods:Hyaluronidase was injected through facial artery in 13 patients who were diagnosed with vascular embolism after facial injection from January 2019 to May 2020. The facial artery was punctured with 22-gauge arterial blood collection needle or 19/23-gauge disposable venous infusion needle. The angle between the needle body and the skin varies depending on the patients’ weight, ranged 30°-45°. The needle was advanced slowly and pushed forward by 2-3 mm when blood backflow appeared in the needle core. After confirming the successful puncture of the facial artery, 0.5-1.5 ml hyaluronidase was slowly injected into the facial artery. The time of skin relaxation, tenderness relief, ulcer healing and wound recovery were observed. The pigmentation was observed and the Vancouver Scar Scale (VSS) was used to score the scars after 3-12 months.Results:A toal of 13 patients with vascular complications of hyaluronidase filler were retrospectively reviewed. The patients were 18-45-year-old(mean age, 35 years) and received hyaluronidase filler at private clinics. There were 12 women and 1 man. The time from onset to visit was 14 h to 4 d, with an average time of 2.5 d. Hyaluronidase was most commonly injected into the nasolabial folds (54%, 7 of 13). The second-ranked area is the nasalroot (23%, 3 of 13). These patients had skin swelling, necrosis, ecchymosis or black scabs during or after hyaluronidase injection. Some patients showed skin lesions combined with oral ulcer. After percutaneous facial arterial hyaluronidase injection, the local skin tissue injuries of the 13 patients were improved in time. The time of skin relaxation was (0.77±0.25) d, the time of tenderness relief was (1.23±0.64) d, the time of ulcer healing was (3.14±0.64) d and the time of wound recovery was (5.85±0.86) d. Patients were followed up for 3-12 months, with an average of 7 months. One patient had slight scar (VSS score of 1), two patients had only mild pigmentation (VSS score of 0), and the other ten patients had no scar and pigmentation (VSS score of 0).Conclusions:It is effective to improve local microcirculation and reduce skin tissue injury after percutaneous facial artery hyaluronidase injection in the treatment of skin injury caused by facial filler injection.

Objective:To screen and identify differentially expressed long non-coding RNA (lncRNAs) in peripheral blood of children with sepsis, and to explore the role of lncRNAs in the pathogenesis of sepsis in children.Methods:The peripheral blood samples of 3 children with sepsis admitted to the ICU of Children′s Hospital of Capital Institute of Pediatrics from January to December 2016 and 3 healthy children who underwent physical examination in our hospital during the same period were selected, and the differential expression profiles of lncRNAs and mRNAs were screened by lncRNAs sequencing technology.The target genes of differentially expressed lncRNAs were predicted and the relationship pairs of lncRNA-mRNA related to F 1F O-ATPase activity were constructed according to the results of GO analysis.Further increasing the sample size, we verified the expression of some F 1F O-ATPase activity-related mRNAs and lncRNAs which were differentially expressed in the screening results by real-time fluorescent quantitative polymerase chain reaction(qRT-PCR). Results:Sequencing results showed that there were 252 lncRNAs with significant differential expression in peripheral blood of children with sepsis compared with healthy children, of which 86 were up-regulated and 166 were down-regulated; meanwhile, there were 2 652 mRNAs with significant differential expression, of which 955 were up-regulated and 1 697 were down-regulated.The results of qRT-PCR showed that the expression of lncRNA ENST00000621933.1, ENST00000616950.1 and ENST00000595748.1 in peripheral blood of children with sepsis increased( P<0.05), while the expression of MT-ATP8, ATP5E and ENST00000624705.1, ENST00000615535.1 in peripheral blood of children with sepsis decreased( P<0.05), which was consistent with the sequencing results. Conclusion:lncRNAs are differentially expressed in peripheral blood of children with sepsis compared with healthy children.The expression levels of lncRNA ENST00000621933.1, ENST00000616950.1, ENST00000595748.1, ENST00000624705.1 and ENST00000615535.1 which their target genes are MT-ATP8 and ATP5E may be related to the development of sepsis in children.

Objective:To investigate the expression of oxidative stress and related pathway factors in fibroblasts from human hypertrophic scar and normal skin tissue.Methods:Select and collect the scar tissue and normal full-thickness skin tissue of the donor area from 8 hypertrophic scar patients who were treated with surgical resection at the Department of Burns and Plastic Surgery, the Fourth Medical Center of Chinese PLA General Hospital from June 2019 to July 2020, extract and culture primary fibroblasts by weaving method, subculture them, and perform the following experiments when the cells were subcultured to the third generation: (1)Detect the proliferation ability of hypertrophic scar fibroblasts(HSF)and normal skin fibroblasts(NSF)with CCK-8 method.(2)Detect the number of reactive oxygen species in HSF and NSF by flow cytometry.(3)Colorimetric method for detecting relative activity of superoxide dismutase(SOD)in two kinds of cells.(4)Detect the NQO1 gene expression in two kinds of cells with RT-PCR.(5)Detect the Nrf2 gene expression in two kinds of cells with RT-PCR.(6)Detects Nrf2 and Bcl2 protein content in two kinds of cells with Western blotting.(7)Detect the expression and distribution of Nrf2 in cells with cellular immunohistochemical staining. The results were analyzed by SPSS 25.0 statistical software, independent sample t test was performed, and P<0.05 was considered as the difference was statistically significant. Results:(1) Compared with NSF, the number of the two kinds of cells is statistically different from the third day (The statistical results from the third day to the seventh day were as follows: t=2.631, P=0.039; t=7.025, P<0.001; t=5.031, P<0.001; t=4.241, P=0.002; t=6.525, P=0.003). (2) The reactive oxygen species content in HSF was 75.39%±3.06%, which was significantly higher than 45.36%±3.66% in the NSF group ( t=-17.804, P<0.001). (3) The relative activity of SOD of the HSF group was (50.76±0.52) U/ml, which is higher than that of the NSF group (42.76±1.35) U/ml ( t=15.674, P<0.001). (4) The relative expression of NQO1 mRNA in HSF was 0.859±0.076, which was lower than that in the NSF group at 1.595±0.181 ( t=3.763, P=0.020). (5) The relative expression of Nrf2 mRNA in HSF was 0.590±0.055, which was lower than that in the NSF group at 1.595±0.146 ( t=6.445, P=0.003). (6) The relative expression of Nrf2 protein of HSF was 0.314±0.035, which was significantly lower than 0.912±0.039 in the NSF group ( t=22.554, P<0.001). The relative expression of Bcl2 protein of HSF was 0.466±0.020, which was significantly lower than 0.734±0.066 in the NSF group ( t=7.780, P<0.001). (7) Nrf2 protein express in NSF and HSF cells, and protein expression was found in both nucleus and cytoplasm. Conclusions:HSF has a high degree of oxidative stress, which may be due to some reasons that reduce the content of Nrf2, resultsing in a decrease in the expression of various antioxidant enzymes, and ultimately leading to the accumulation of reactive oxygen species. Compared with NSF, HSF has stronger proliferation and apoptosis abilities, and reactive oxygen species can cause cell apoptosis, indicating that oxidative stress may be one of the pathogenesis of hypertrophic scars.

Objective:To investigate the expression of oxidative stress and related pathway factors in fibroblasts from human hypertrophic scar and normal skin tissue.Methods:Select and collect the scar tissue and normal full-thickness skin tissue of the donor area from 8 hypertrophic scar patients who were treated with surgical resection at the Department of Burns and Plastic Surgery, the Fourth Medical Center of Chinese PLA General Hospital from June 2019 to July 2020, extract and culture primary fibroblasts by weaving method, subculture them, and perform the following experiments when the cells were subcultured to the third generation: (1)Detect the proliferation ability of hypertrophic scar fibroblasts(HSF)and normal skin fibroblasts(NSF)with CCK-8 method.(2)Detect the number of reactive oxygen species in HSF and NSF by flow cytometry.(3)Colorimetric method for detecting relative activity of superoxide dismutase(SOD)in two kinds of cells.(4)Detect the NQO1 gene expression in two kinds of cells with RT-PCR.(5)Detect the Nrf2 gene expression in two kinds of cells with RT-PCR.(6)Detects Nrf2 and Bcl2 protein content in two kinds of cells with Western blotting.(7)Detect the expression and distribution of Nrf2 in cells with cellular immunohistochemical staining. The results were analyzed by SPSS 25.0 statistical software, independent sample t test was performed, and P<0.05 was considered as the difference was statistically significant. Results:(1) Compared with NSF, the number of the two kinds of cells is statistically different from the third day (The statistical results from the third day to the seventh day were as follows: t=2.631, P=0.039; t=7.025, P<0.001; t=5.031, P<0.001; t=4.241, P=0.002; t=6.525, P=0.003). (2) The reactive oxygen species content in HSF was 75.39%±3.06%, which was significantly higher than 45.36%±3.66% in the NSF group ( t=-17.804, P<0.001). (3) The relative activity of SOD of the HSF group was (50.76±0.52) U/ml, which is higher than that of the NSF group (42.76±1.35) U/ml ( t=15.674, P<0.001). (4) The relative expression of NQO1 mRNA in HSF was 0.859±0.076, which was lower than that in the NSF group at 1.595±0.181 ( t=3.763, P=0.020). (5) The relative expression of Nrf2 mRNA in HSF was 0.590±0.055, which was lower than that in the NSF group at 1.595±0.146 ( t=6.445, P=0.003). (6) The relative expression of Nrf2 protein of HSF was 0.314±0.035, which was significantly lower than 0.912±0.039 in the NSF group ( t=22.554, P<0.001). The relative expression of Bcl2 protein of HSF was 0.466±0.020, which was significantly lower than 0.734±0.066 in the NSF group ( t=7.780, P<0.001). (7) Nrf2 protein express in NSF and HSF cells, and protein expression was found in both nucleus and cytoplasm. Conclusions:HSF has a high degree of oxidative stress, which may be due to some reasons that reduce the content of Nrf2, resultsing in a decrease in the expression of various antioxidant enzymes, and ultimately leading to the accumulation of reactive oxygen species. Compared with NSF, HSF has stronger proliferation and apoptosis abilities, and reactive oxygen species can cause cell apoptosis, indicating that oxidative stress may be one of the pathogenesis of hypertrophic scars.

Objective:To explore the psychology and influencing factors of decision-making for giving up breast-conserving and choosing unilateral mastectomy among breast cancer patients with feasible breast-conserving surgery.Methods:From September 2019 to February 2020, we selected 18 breast cancer patients with breast-conserving indications and finally choosing unilateral mastectomy in Breast Surgery at Shanxi Bethune Hospital as the study objects by purposive sampling. The qualitative research was used to carry out semi-structured in-depth interview, and the data was organized and analyzed.Results:From four aspects, the patient factor, family factor, medical and nursing staff factor as well as social factor, a total of 8 themes that affected decision-making were summarized, including that patients were under financial pressure, fatigue, and had cognition bias and expected sadness; family members concealed the condition, and lack of family members and patients participation in decision-making; inadequate health education by medical and nursing staff; network information needed to be discussed, and misdirection of peer experience.Conclusions:Decision-making for surgical methods of breast cancer patients is influenced by many factors. Medical and nursing staff should use the existing factors as an entry point to construct intervention measures so as to promote patient participation in clinical decision-making.